焦脱镁叶绿酸甲酯介导光动力抑制人乳腺癌细胞MCF-7迁移

Pyropheophorbide-a methyl ester-mediated photodynamic therapy inhibits migration of human breast cancer MCF-7 cells

目的探讨光敏剂焦脱镁叶绿酸甲酯(pyropheophorbide-a methyl ester,MPPa)介导光动力治疗(photodynamic therapy, PDT)对人乳腺癌细胞MCF-7迁移的影响及其可能的作用机制。方法将处于对数期生长的人乳腺癌细胞MCF-7随机分为对照组、单药组(仅MPPa处理)、单光组(仅光照处理)和MPPa-PDT组(MPPa和光照处理),使用不同浓度MPPa(0、0.5、1、2、4μmol·L^(-1))及不同光照强度(0、0.9、1.8、2.7、3.6 J/cm^2)干预MCF-7细胞,CCK-8法检测细胞活性;DCFH-DA染色检测细胞内活性氧(reactive oxygen species, ROS)的产生;Transwell、细胞划痕实验观察细胞迁移能力;Western blot检测E-cadherin、PTEN、Akt、p-Akt蛋白的表达变化。结果与对照组、单药组和单光组相比, MPPa-PDT能抑制MCF-7细胞活性,且抑制作用呈剂量依赖性;DCFH-DA染色显示,MPPa-PDT组细胞内ROS产生明显高于各对照组;Transwell及划痕实验结果表明,MPPa-PDT组细胞迁移能力较各对照组明显降低;Western blot结果表明,MPPa-PDT组E-cadherin、PTEN蛋白表达升高,p-Akt蛋白表达降低。结论 MPPa介导光动力可抑制人乳腺癌细胞MCF-7迁移能力,其机制可能涉及PTEN/Akt通路。

Aim To investigate the effect of pyropheophorbide-a methyl ester-mediated photodynamic therapy(MPPa-PDT)on migration in human breast cancer MCF-7cells as well as its possible molecular mechanism.Methods MCF-7cells were randomly divided into control group,MPPa group,light group and MPPa-PDT group.Cells were co-cultured with MPPa at different concentrations(0,0.5,1,2,4μmol·L^-1)and exposed to light of different energy intensities(0,0.9,1.8,2.7,3.6J/cm^2).Cell viability was measured by CCK-8;production of ROS was detected by DCFH-DA staining;cell migration was mesured by Transwell migration assay and wound healing assay;the expressions of E-cadherin,PTEN,Akt,p-Akt were measured by Western blot.Results Compared with control group,MPPa group and light group,MPPa-PDT could significantly inhibit cell viability of MCF-7cells in an energy-dependent manner.DCFH-DA staining suggested ROS in MPPa-PDT group was higher than that in the other three control groups;Transwell assay and wound healing assay showed cell migration in MPPa-PDT group was significantly inhibited;Western blot analysis indicated that the expression of E-cadherin,PTEN in MPPa-PDT group increased,while p-Akt decreased.Conclusions MPPa-PDT inhibits MCF-7cells’migration,and PTEN/Akt signal pathway may be involved in the process.